206 results on '"Tweardy DJ"'
Search Results
2. "Personalizing" academic medicine: opportunities and challenges in implementing genomic profiling.
- Author
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Tweardy DJ, Belmont JW, Tweardy, David J, and Belmont, John W
- Abstract
BCM faculty members spearheaded the development of a first-generation Personal Genome Profile (Baylor PGP) assay to assist physicians in diagnosing and managing patients in this new era of medicine. The principles that guided the design and implementation of the Baylor PGP were high quality, robustness, low expense, flexibility, practical clinical utility, and the ability to facilitate broad areas of clinical research. The most distinctive feature of the approach taken is an emphasis on extensive screening for rare disease-causing mutations rather than common risk-increasing polymorphisms. Because these variants have large direct effects, the ability to screen for them inexpensively could have a major immediate clinical impact in disease diagnosis, carrier detection, presymptomatic detection of late onset disease, and even prenatal diagnosis. In addition to creating a counseling tool for individual "consumers," this system will fit into the established medical record and be used by physicians involved in direct patient care. This article describes an overall framework for clinical diagnostic array genotyping and the available technologies, as well as highlights the opportunities and challenges for implementation. [ABSTRACT FROM AUTHOR]
- Published
- 2009
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3. Interleukin-6 treatment reverses apoptosis and blunts susceptibility to intraperitoneal bacterial challenge following hemorrhagic shock.
- Author
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Arikan AA, Mastrangelo MA, Tweardy DJ, Arikan, Ayse Akcan, Yu, Bi, Mastrangelo, Mary-Ann, and Tweardy, David J
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- 2006
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4. Single nucleotide polymorphisms in genes for 2'-5'-oligoadenylate synthetase and RNase L inpatients hospitalized with West Nile virus infection.
- Author
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Yakub I, Lillibridge KM, Moran A, Gonzalez OY, Belmont J, Gibbs RA, Tweardy DJ, Yakub, Imtiaz, Lillibridge, Kristy M, Moran, Ana, Gonzalez, Omar Y, Belmont, John, Gibbs, Richard A, and Tweardy, David J
- Abstract
Background: Infection with the flavivirus West Nile virus (WNV) is a growing problem across the United States, where there is a case-fatality rate of 15%-29% in individuals >70 years old and no consistently effective treatment. Susceptibility to WNV disease in inbred strains of mice was mapped to a nonsense mutation in the gene encoding the 1b isoform of 2'-5'-oligoadenylate synthetase (OAS), a member of the OAS/RNase L system of innate viral resistance. Genetic susceptibility to severe WNV disease in humans has not been determined.Methods: We sequenced each exon within all OAS and RNASEL genes in 33 individuals hospitalized with WNV infection in Houston to assess if there is a defect in this system in patients with severe WNV disease.Results: Sequencing did not reveal any insertions, deletions, or nonsense mutations in any OAS or RNASEL gene. However, comparison of the exonic sequences between case patients and control subjects identified 23 single nucleotide polymorphisms (SNPs), including a synonymous SNP in OASL exon 2 (rs3213545), in which the reference allele occurred at a higher frequency in case patients (P < .004).Conclusion: Because the reference allele contains a splice enhancer site, our finding suggests that the RNA transcripts generated from this allele may undergo increased splicing, which results in a dominant-negative OASL isozyme similar to the nonsense/truncation mutant form of Oas1b in mice. [ABSTRACT FROM AUTHOR]- Published
- 2005
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5. Lethal human neutropenic entercolitis caused by Clostridium chauvoei in the United States: tip of the iceberg?
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Weatherhead JE and Tweardy DJ
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- 2012
6. Substance P signaling contributes to granuloma formation in Taenia crassiceps infection, a murine model of cysticercosis.
- Author
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Garza A, Tweardy DJ, Weinstock J, Viswanathan B, and Robinson P
- Abstract
Cysticercosis is an infection with larval cysts of the cestode Taenia solium. Through pathways that are incompletely understood, dying parasites initiate a granulomatous reaction that, in the brain, causes seizures. Substance P (SP), a neuropeptide involved in pain-transmission, contributes to inflammation and previously was detected in granulomas associated with dead T. crassiceps cysts. To determine if SP contributes to granuloma formation, we measured granuloma-size and levels of IL-1beta, TNF-alpha, and IL-6 within granulomas in T. crassiceps-infected wild type (WT) mice and mice deficient in SP-precursor (SPP) or the SP-receptor (neurokinin 1, NK1). Granuloma volumes of infected SPP- and NK1-knockout mice were reduced by 31 and 36%, respectively, compared to WT mice (P < .05 for both) and produced up to 5-fold less IL-1beta, TNF-alpha, and IL-6 protein. Thus, SP signaling contributes to granuloma development and proinflammatory cytokine production in T. crassiceps infection and suggests a potential role for this mediator in human cystercercosis. [ABSTRACT FROM AUTHOR]
- Published
- 2010
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7. Therapeutic Potential of a Small-Molecule STAT3 Inhibitor in a Mouse Model of Colitis.
- Author
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Robinson P, Montoya K, Magness E, Rodriguez E, Villalobos V, Engineer N, Yang P, Bharadwaj U, Eckols TK, and Tweardy DJ
- Abstract
Background and Aims: Inflammatory bowel disease (IBD) predisposes to colorectal cancer (CRC). In the current studies, we used the dextran sodium sulfate (DSS) murine model of colitis, which is widely used in preclinical studies, to determine the contribution of STAT3 to IBD. STAT3 has two isoforms: (STAT3 α; which has pro-inflammatory and anti-apoptotic functions, and STAT3β; which attenuates the effects of STAT3α). In the current study, we determined the contribution of STAT3 to IBD across all tissues by examining DSS-induced colitis in mice that express only STAT3α and in mice treated with TTI-101, a direct small-molecule inhibitor of both isoforms of STAT3., Methods: We examined mortality, weight loss, rectal bleeding, diarrhea, colon shortening, apoptosis of colonic CD4+ T-cells, and colon infiltration with IL-17-producing cells following 7-day administration of DSS (5%) to transgenic STAT3α knock-in (STAT3β-deficient; ΔβΔβ) mice and wild-type (WT) littermate cage control mice. We also examined the effect of TTI-101 on these endpoints in DSS-induced colitis in WT mice., Results: Each of the clinical manifestations of DSS-induced colitis examined was exacerbated in ΔβΔβ transgenic versus cage-control WT mice. Importantly, TTI-101 treatment of DSS-administered WT mice led to complete attenuation of each of the clinical manifestations and also led to increased apoptosis of colonic CD4+ T cells, reduced colon infiltration with IL-17-producing cells, and down-modulation of colon mRNA levels of STAT3-upregulated genes involved in inflammation, apoptosis resistance, and colorectal cancer metastases., Conclusions: Thus, small-molecule targeting of STAT3 may be of benefit in treating IBD and preventing IBD-associated colorectal cancer.
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- 2023
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8. Aberrant function of pathogenic STAT3 mutant proteins is linked to altered stability of monomers and homodimers.
- Author
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Kasembeli MM, Kaparos E, Bharadwaj U, Allaw A, Khouri A, Acot B, and Tweardy DJ
- Subjects
- Mutation, DNA metabolism, Tyrosine genetics, STAT3 Transcription Factor genetics, STAT3 Transcription Factor metabolism, src Homology Domains genetics
- Abstract
STAT3 mutations, predominantly in the DNA-binding domain (DBD) and Src-homology 2 domain (SH2D), cause rare cases of immunodeficiency, malignancy, and autoimmunity. The exact mechanisms by which these mutations abrogate or enhance STAT3 function are not completely understood. Here, we examined how loss-of-function (LOF) and gain-of-function (GOF) STAT3 mutations within the DBD and SH2D affect monomer and homodimer protein stability as well as their effect on key STAT3 activation events, including recruitment to phosphotyrosine (pY) sites within peptide hormone receptors, tyrosine phosphorylation at Y705, dimerization, nuclear translocation, and DNA binding. The DBD LOF mutants showed reduced DNA binding when homodimerized, whereas the DBD GOF mutants showed increased DNA binding. DBD LOF and GOF mutants showed minimal changes in other STAT3 functions or in monomer or homodimer protein stability. However, SH2D LOF mutants demonstrated reduced conformational stability as either monomers or homodimers, leading to decreased pY-peptide recruitment, tyrosine phosphorylation, dimerization, nuclear localization, and DNA binding. In contrast, cancer-causing SH2D GOF mutants showed increased STAT3 homodimer stability, which increased their DNA binding. Of note, a small-molecule inhibitor of STAT3 that targets the tyrosine phosphopeptide-binding pocket within the STAT3 SH2D potently inhibited cell proliferation driven by STAT3 SH2D GOF mutants. These findings indicate that the stability of STAT3 protein monomer and homodimer is critical for the pathogenesis of diseases caused by SH2D LOF and GOF mutations and suggest that agents that modulate STAT3 monomer and/or homodimer protein stability may have therapeutic value in diseases caused by these mutations., (© 2023 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)
- Published
- 2023
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9. Genetic and Small-Molecule Modulation of Stat3 in a Mouse Model of Crohn's Disease.
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Robinson P, Magness E, Montoya K, Engineer N, Eckols TK, Rodriguez E, and Tweardy DJ
- Abstract
Crohn's disease (CD), is an inflammatory bowel disease that can affect any part of the gastro-intestinal tract (GI) and is associated with an increased risk of gastro-intestinal cancer. In the current study, we determined the role of genetic and small-molecule modulation of STAT3 in a mouse model of CD. STAT3 has 2 isoforms (α, β) which are expressed in most cells in a 4:1 ratio (α: β). STAT3α has pro-inflammatory and anti-apoptotic functions, while STAT3β has contrasting roles. We used an animal model of CD consisting of intrarectal administration of 2,4,6-trinitrobenzene sulfonic acid and examined the severity of CD in transgenic-mice that express only STAT3α (∆
β /∆β ), as well as in wild-type (WT) mice administered TTI-101 (formerly C188-9), a small molecule STAT3 inhibitor. We determined that clinical manifestations of CD, such as mortality, rectal-bleeding, colonic bleeding, diarrhea, and colon shortening, were exacerbated in ∆β /∆β transgenic versus cage-control WT mice, while they were markedly decreased by TTI-101 treatment of WT mice. TTI-101 treatment also increased apoptosis of pathogenic CD4+ T cells and reduced colon levels of IL-17-positive cells. Our results indicate that STAT3 contributes to CD and that targeting of STAT3 with TTI-101 may be a useful approach to treating CD.- Published
- 2022
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10. Spatial molecular and cellular determinants of STAT3 activation in liver fibrosis progression in non-alcoholic fatty liver disease.
- Author
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Jiao J, Sanchez JI, Saldarriaga OA, Solis LM, Tweardy DJ, Maru DM, Stevenson HL, and Beretta L
- Abstract
Background & Aims: The prevalence of non-alcoholic fatty liver disease (NAFLD) and its severe form, non-alcoholic steatohepatitis (NASH), is increasing. Individuals with NASH often develop liver fibrosis and advanced liver fibrosis is the main determinant of mortality in individuals with NASH. We and others have reported that STAT3 contributes to liver fibrosis and hepatocellular carcinoma in mice., Methods: Here, we explored whether STAT3 activation in hepatocyte and non-hepatocyte areas, measured by phospho-STAT3 (pSTAT3), is associated with liver fibrosis progression in 133 patients with NAFLD. We further characterized the molecular and cellular determinants of STAT3 activation by integrating spatial distribution and transcriptomic changes in fibrotic NAFLD livers.Results: pSTAT3 scores in non-hepatocyte areas progressively increased with fibrosis severity (r = 0.53, p <0.001). Correlation analyses between pSTAT3 scores and expression of 1,540 immune- and cancer-associated genes revealed a large effect of STAT3 activation on gene expression changes in non-hepatocyte areas and confirmed a major role for STAT3 activation in fibrogenesis. Digital spatial transcriptomic profiling was also performed on 13 regions selected in hepatocyte and non-hepatocyte areas from four NAFLD liver biopsies with advanced fibrosis, using a customized panel of markers including pSTAT3, PanCK+CK8/18, and CD45. The regions were further segmented based on positive or negative pSTAT3 staining. Cell deconvolution analysis revealed that activated STAT3 was enriched in hepatic progenitor cells (HPCs) and sinusoidal endothelial cells. Regression of liver fibrosis upon STAT3 inhibition in mice with NASH resulted in a reduction of HPCs, demonstrating a direct role for STAT3 in HPC expansion., Conclusion: Increased understanding of the spatial dependence of STAT3 signaling in NASH and liver fibrosis progression could lead to novel targeted treatment approaches., Impact and Implications: Advanced liver fibrosis is the main determinant of mortality in patients with NASH. This study showed using liver biopsies from 133 patients with NAFLD, that STAT3 activation in non-hepatocyte areas is strongly associated with fibrosis severity, inflammation, and progression to NASH. STAT3 activation was enriched in hepatic progenitor cells (HPCs) and sinusoidal endothelial cells (SECs), as determined by innovative technologies interrogating the spatial distribution of pSTAT3. Finally, STAT3 inhibition in mice resulted in reduced liver fibrosis and depletion of HPCs, suggesting that STAT3 activation in HPCs contributes to their expansion and fibrogenesis in NAFLD., Competing Interests: The authors declare no conflicts of interest that pertain to this work. Please refer to the accompanying ICMJE disclosure forms for further details., (© 2022 The Author(s).)
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- 2022
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11. Corrigendum to "TTI-101: A competitive inhibitor of STAT3 that spares oxidative phosphorylation and reverses mechanical allodynia in mouse models of neuropathic pain" [Biochem. Pharmacol. 192 (2021) 114688].
- Author
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Kasembeli MM, Singhmar P, Ma J, Edralin J, Tang Y, Adams C 3rd, Heijnen CJ, Kavelaars A, and Tweardy DJ
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- 2022
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12. DRUGGING "UNDRUGGABLE" DISEASE-CAUSING PROTEINS: FOCUS ON SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION (STAT) 3.
- Author
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Tweardy DJ
- Subjects
- Humans, Ligands, Pharmaceutical Preparations, Naphthols, Sulfonamides
- Abstract
Signal transducer and activator of transcription (STAT) 3 has been assigned to the group of "undruggable" disease-causing proteins, despite its containing a Src-homology (SH) 2 domain, a potential Achilles' heel that has eluded successful targeting by academic and pharmaceutical groups over the past 30 years. Based on mutational and modeling studies, our group developed a unique virtual ligand screening strategy targeting the STAT3 SH2 domain that was coupled to robust biochemical and cellular assays and structure-based medicinal chemistry and led to the identification of TTI-101. TTI-101 represents one of the most advanced, direct, small-molecule inhibitors of an SH2 domain-containing, disease-causing protein in clinical development. TTI-101 is currently being evaluated in a Phase 1 study to determine safety and tolerability in addition to pharmacodynamic effects and efficacy in patients with advanced solid tumors., Competing Interests: Potential Conflicts of Interest: David J. Tweardy co-founded and holds equity in Tvardi Therapeutics. The intellectual property used in the research supporting this publication was created by Dr. Tweardy and licensed to Tvardi Therapeutics, Inc. Dr. Tweardy also receives compensation from Tvardi as a member of its scientific advisory board., (© 2022 The American Clinical and Climatological Association.)
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- 2022
13. TTI-101: A competitive inhibitor of STAT3 that spares oxidative phosphorylation and reverses mechanical allodynia in mouse models of neuropathic pain.
- Author
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Kasembeli MM, Singhmar P, Ma J, Edralin J, Tang Y, Adams C 3rd, Heijnen CJ, Kavelaars A, and Tweardy DJ
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- Animals, Cell Line, Tumor, Disease Models, Animal, Female, Humans, Hyperalgesia metabolism, Male, Mice, Mice, Inbred C57BL, Naphthols pharmacology, Neuralgia metabolism, STAT3 Transcription Factor metabolism, Sulfonamides pharmacology, Hyperalgesia drug therapy, Naphthols therapeutic use, Neuralgia drug therapy, Oxidative Phosphorylation drug effects, STAT3 Transcription Factor antagonists & inhibitors, Sulfonamides therapeutic use, Touch
- Abstract
Signal Transducer and Activator of Transcription (STAT) 3 emerged rapidly as a high-value target for treatment of cancer. However, small-molecule STAT3 inhibitors have been slow to enter the clinic due, in part, to serious adverse events (SAE), including lactic acidosis and peripheral neuropathy, which have been attributed to inhibition of STAT3's mitochondrial function. Our group developed TTI-101, a competitive inhibitor of STAT3 that targets the receptor pY705-peptide binding site within the Src homology 2 (SH2) domain to block its recruitment and activation. TTI-101 has shown target engagement, no toxicity, and evidence of clinical benefit in a Phase I study in patients with solid tumors. Here we report that TTI-101 did not affect mitochondrial function, nor did it cause STAT3 aggregation, chemically modify STAT3 or cause neuropathic pain. Instead, TTI-101 unexpectedly suppressed neuropathic pain induced by chemotherapy or in a spared nerve injury model. Thus, in addition to its direct anti-tumor effect, TTI-101 may be of benefit when administered to cancer patients at risk of developing chemotherapy-induced peripheral neuropathy (CIPN)., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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14. Signal Transducer and Activator of Transcription-3 Modulation of Cardiac Pathology in Chronic Chagasic Cardiomyopathy.
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Hoffman KA, Villar MJ, Poveda C, Bottazzi ME, Hotez PJ, Tweardy DJ, and Jones KM
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- Animals, Heart, Mice, Chagas Cardiomyopathy drug therapy, Chagas Disease drug therapy, STAT3 Transcription Factor antagonists & inhibitors, Trypanosoma cruzi
- Abstract
Chronic Chagasic cardiomyopathy (CCC) is a severe clinical manifestation that develops in 30%-40% of individuals chronically infected with the protozoal parasite Trypanosoma cruzi and is thus an important public health problem. Parasite persistence during chronic infection drives pathologic changes in the heart, including myocardial inflammation and progressive fibrosis, that contribute to clinical disease. Clinical manifestations of CCC span a range of symptoms, including cardiac arrhythmias, thromboembolic disease, dilated cardiomyopathy, and heart failure. This study aimed to investigate the role of signal transducer and activator of transcription-3 (STAT3) in cardiac pathology in a mouse model of CCC. STAT3 is a known cellular mediator of collagen deposition and fibrosis. Mice were infected with T. cruzi and then treated daily from 70 to 91 days post infection (DPI) with TTI-101, a small molecule inhibitor of STAT3; benznidazole; a combination of benznidazole and TTI-101; or vehicle alone. Cardiac function was evaluated at the beginning and end of treatment by echocardiography. By the end of treatment, STAT3 inhibition with TTI-101 eliminated cardiac fibrosis and fibrosis biomarkers but increased cardiac inflammation; serum levels of interleukin-6 (IL-6), and IFN- γ ; cardiac gene expression of STAT1 and nuclear factor-κB (NF-κB); and upregulation of IL-6 and Type I and Type II IFN responses. Concurrently, decreased heart function was measured by echocardiography and myocardial strain. These results indicate that STAT3 plays a critical role in the cardiac inflammatory-fibrotic axis during CCC., Competing Interests: DT is the inventor of several patents concerning TTI-101 that are owned by Baylor College of Medicine and licensed to Tvardi Therapeutics, Inc.; DT has ownership of this company’s stock. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Hoffman, Villar, Poveda, Bottazzi, Hotez, Tweardy and Jones.)
- Published
- 2021
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15. Novel STAT3 small-molecule inhibitors identified by structure-based virtual ligand screening incorporating SH2 domain flexibility.
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Kong R, Bharadwaj U, Eckols TK, Kolosov M, Wu H, Cruz-Pavlovich FJS, Shaw A, Ifelayo OI, Zhao H, Kasembeli MM, Wong STC, and Tweardy DJ
- Subjects
- Alkylation, Binding Sites drug effects, Blotting, Western, Cell Line, Tumor drug effects, Gas Chromatography-Mass Spectrometry, Humans, Ligands, Molecular Docking Simulation, Protein Structure, Tertiary, STAT3 Transcription Factor chemistry, STAT3 Transcription Factor genetics, Structure-Activity Relationship, Surface Plasmon Resonance, Drug Evaluation, Preclinical methods, STAT3 Transcription Factor antagonists & inhibitors, src Homology Domains drug effects
- Abstract
Efforts to develop STAT3 inhibitors have focused on its SH2 domain starting with short phosphotyrosylated peptides based on STAT3 binding motifs, e.g. pY
905 LPQTV within gp130. Despite binding to STAT3 with high affinity, issues regarding stability, bioavailability, and membrane permeability of these peptides, as well as peptidomimetics such as CJ-887, have limited their further clinical development and led to interest in small-molecule inhibitors. Some small molecule STAT3 inhibitors, identified using structure-based virtual ligand screening (SB-VLS); while having favorable drug-like properties, suffer from weak binding affinities, possibly due to the high flexibility of the target domain. We conducted molecular dynamic (MD) simulations of the SH2 domain in complex with CJ-887, and used an averaged structure from this MD trajectory as an "induced-active site" receptor model for SB-VLS of 110,000 compounds within the SPEC database. Screening was followed by re-docking and re-scoring of the top 30% of hits, selection for hit compounds that directly interact with pY + 0 binding pocket residues R609 and S613, and testing for STAT3 targeting in vitro, which identified two lead hits with good activity and favorable drug-like properties. Unlike most small-molecule STAT3 inhibitors previously identified, which contain negatively-charged moieties that mediate binding to the pY + 0 binding pocket, these compounds are uncharged and likely will serve as better candidates for anti-STAT3 drug development. IMPLICATIONS: SB-VLS, using an averaged structure from molecular dynamics (MD) simulations of STAT3 SH2 domain in a complex with CJ-887, a known peptidomimetic binder, identify two highly potent, neutral, low-molecular weight STAT3-inhibitors with favorable drug-like properties., (Published by Elsevier Ltd.)- Published
- 2021
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16. Pharmacokinetics and pharmacodynamics of TTI-101, a STAT3 inhibitor that blocks muscle proteolysis in rats with chronic kidney disease.
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Zhang L, Wang Y, Dong Y, Chen Z, Eckols TK, Kasembeli MM, Tweardy DJ, and Mitch WE
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- Animals, Body Weight drug effects, Chromatography, Liquid, Disease Models, Animal, Enzyme Inhibitors pharmacokinetics, Hand Strength, Muscle, Skeletal metabolism, Naphthols pharmacokinetics, Rats, Sulfonamides pharmacokinetics, Tandem Mass Spectrometry, Enzyme Inhibitors pharmacology, Muscle, Skeletal drug effects, Naphthols pharmacology, Proteolysis drug effects, Renal Insufficiency, Chronic metabolism, STAT3 Transcription Factor antagonists & inhibitors, Sulfonamides pharmacology
- Abstract
Loss of muscle proteins increases the morbidity and mortality of patients with chronic kidney disease (CKD), and there are no reliable preventive treatments. We uncovered a STAT3/CCAAT-enhancer-binding protein-δ to myostatin signaling pathway that activates muscle protein degradation in mice with CKD or cancer; we also identified a small-molecule inhibitor of STAT3 (TTI-101) that blocks this pathway. To evaluate TTI-101 as a treatment for CKD-induced cachexia, we measured TTI-101 pharmacokinetics and pharmacodynamics in control and CKD rats that were orally administered TTI-101or its diluent. The following two groups of gavage-fed rats were studied: sham-operated control rats and CKD rats. Plasma was collected serially (0, 0.25, 0.5, 1, 2, 4, 8, and 24 h) following TTI-101 administration (at oral doses of 0, 10, 30, or 100 mg/kg). Plasma levels of TTI-101 were measured by LC-MS/MS, and pharmacokinetic results were analyzed with the PKSolver program. Plasma TTI-101 levels increased linearly with doses; the maximum plasma concentrations and time to maximal plasma levels (~1 h) were similar in sham-operated control rats and CKD rats. Notably, gavage treatment of TTI-101 for 3 days produced TTI-101 muscle levels in sham control rats and CKD rats that were not significantly different. CKD rats that received TTI-101 for 7 days had suppression of activated STAT3 and improved muscle grip strength; there also was a trend for increasing body and muscle weights. TTI-101 was tolerated at doses of 100 mg·kg
-1 ·day-1 for 7 days. These results with TTI-101 in rats warrant its development as a treatment for cachexia in humans.- Published
- 2020
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17. Targeting STAT3 anti-apoptosis pathways with organic and hybrid organic-inorganic inhibitors.
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Minus MB, Wang H, Munoz JO, Stevens AM, Mangubat-Medina AE, Krueger MJ, Liu W, Kasembeli MM, Cooper JC, Kolosov MI, Tweardy DJ, Redell MS, and Ball ZT
- Subjects
- Animals, Antineoplastic Agents pharmacology, Apoptosis drug effects, Cell Line, Tumor, Drug Screening Assays, Antitumor, Humans, Mice, Models, Molecular, Molecular Targeted Therapy, Neoplasms, Experimental, Oxidation-Reduction, Protein Binding, Protein Kinase Inhibitors pharmacology, STAT3 Transcription Factor genetics, Structure-Activity Relationship, Antineoplastic Agents chemistry, Leukemia, Myeloid, Acute drug therapy, Naphthalenes chemistry, Protein Kinase Inhibitors chemistry, STAT3 Transcription Factor antagonists & inhibitors, Sulfonamides chemistry
- Abstract
Recurrence and drug resistance are major challenges in the treatment of acute myeloid leukemia (AML) that spur efforts to identify new clinical targets and active agents. STAT3 has emerged as a potential target in resistant AML, but inhibiting STAT3 function has proven challenging. This paper describes synthetic studies and biological assays for a naphthalene sulfonamide inhibitor class of molecules that inhibit G-CSF-induced STAT3 phosphorylation in cellulo and induce apoptosis in AML cells. We describe two different approaches to inhibitor design: first, variation of substituents on the naphthalene sulfonamide core allows improvements in anti-STAT activity and creates a more thorough understanding of anti-STAT SAR. Second, a novel approach involving hybrid sulfonamide-rhodium(ii) conjugates tests our ability to use cooperative organic-inorganic binding for drug development, and to use SAR studies to inform metal conjugate design. Both approaches have produced compounds with improved binding potency. In vivo and in cellulo experiments further demonstrate that these approaches can also lead to improved activity in living cells, and that compound 3aa slows disease progression in a xenograft model of AML.
- Published
- 2020
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18. Stat3 activation induces insulin resistance via a muscle-specific E3 ubiquitin ligase Fbxo40.
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Zhang L, Chen Z, Wang Y, Tweardy DJ, and Mitch WE
- Subjects
- Animals, Diet, High-Fat, Insulin metabolism, Insulin Receptor Substrate Proteins metabolism, Mice, Mice, Knockout, Phosphorylation, Proto-Oncogene Proteins c-akt metabolism, STAT3 Transcription Factor genetics, Signal Transduction physiology, F-Box Proteins metabolism, Insulin Resistance physiology, Muscle, Skeletal metabolism, Renal Insufficiency, Chronic metabolism, STAT3 Transcription Factor metabolism
- Abstract
Cellular mechanisms causing insulin resistance (IR) in chronic kidney disease (CKD) are poorly understood. One potential mechanism is that CKD-induced inflammation activates the signal transducer and activator of transcription 3 (Stat3) in muscle. We uncovered increased p-Stat3 in muscles of mice with CKD or mice fed high-fat diet (HFD). Activated Stat3 stimulates the expression of Fbxo40, a muscle-specific E3 ubiquitin ligase that stimulates ubiquitin conjugation leading to degradation of insulin receptor substrate 1 (IRS1). Evidence that Stat3 activates Fbxo40 includes 1 ) potential Stat3 binding sites in Fbxo40 promoters; 2 ) Stat3 binding to the Fbxo40 promoter; and 3 ) constitutively active Stat3 stimulating both Fbxo40 expression and its promoter activity. We found that IL-6 activates Stat3 in myotubes, increasing Fbxo40 expression with reduced IRS1 and p-Akt. Knockdown Fbxo40 using siRNA from myotubes results in higher levels of IRS1 and p-Akt despite the presence of IL-6. We treated mice with a small-molecule inhibitor of Stat3 (TTI-101) and found improved glucose tolerance and insulin signaling in skeletal muscles of mice with CKD or fed an HFD. Finally, we uncovered improved glucose tolerance in mice with muscle-specific Stat3 KO versus results in Stat3
f/f mice in response to the HFD. Thus Stat3 activation in muscle increases IR in mice. Inhibition of Stat3 by TTI-101 could be developed into clinical strategies to improve muscle insulin signaling in inflammation and other catabolic diseases.- Published
- 2020
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19. Targeting Janus Kinases and Signal Transducer and Activator of Transcription 3 to Treat Inflammation, Fibrosis, and Cancer: Rationale, Progress, and Caution.
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Bharadwaj U, Kasembeli MM, Robinson P, and Tweardy DJ
- Subjects
- Animals, Anti-Inflammatory Agents pharmacology, Anti-Inflammatory Agents therapeutic use, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Clinical Trials, Phase II as Topic, Fibrosis metabolism, Humans, Inflammation metabolism, Janus Kinases genetics, Molecular Targeted Therapy, Neoplasms metabolism, Phosphorylation, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors therapeutic use, STAT3 Transcription Factor genetics, Fibrosis drug therapy, Inflammation drug therapy, Janus Kinases antagonists & inhibitors, Janus Kinases metabolism, Neoplasms drug therapy, STAT3 Transcription Factor antagonists & inhibitors, STAT3 Transcription Factor metabolism
- Abstract
Before it was molecularly cloned in 1994, acute-phase response factor or signal transducer and activator of transcription (STAT)3 was the focus of intense research into understanding the mammalian response to injury, particularly the acute-phase response. Although known to be essential for liver production of acute-phase reactant proteins, many of which augment innate immune responses, molecular cloning of acute-phase response factor or STAT3 and the research this enabled helped establish the central function of Janus kinase (JAK) family members in cytokine signaling and identified a multitude of cytokines and peptide hormones, beyond interleukin-6 and its family members, that activate JAKs and STAT3, as well as numerous new programs that their activation drives. Many, like the acute-phase response, are adaptive, whereas several are maladaptive and lead to chronic inflammation and adverse consequences, such as cachexia, fibrosis, organ dysfunction, and cancer. Molecular cloning of STAT3 also enabled the identification of other noncanonical roles for STAT3 in normal physiology, including its contribution to the function of the electron transport chain and oxidative phosphorylation, its basal and stress-related adaptive functions in mitochondria, its function as a scaffold in inflammation-enhanced platelet activation, and its contributions to endothelial permeability and calcium efflux from endoplasmic reticulum. In this review, we will summarize the molecular and cellular biology of JAK/STAT3 signaling and its functions under basal and stress conditions, which are adaptive, and then review maladaptive JAK/STAT3 signaling in animals and humans that lead to disease, as well as recent attempts to modulate them to treat these diseases. In addition, we will discuss how consideration of the noncanonical and stress-related functions of STAT3 cannot be ignored in efforts to target the canonical functions of STAT3, if the goal is to develop drugs that are not only effective but safe. SIGNIFICANCE STATEMENT: Key biological functions of Janus kinase (JAK)/signal transducer and activator of transcription (STAT)3 signaling can be delineated into two broad categories: those essential for normal cell and organ development and those activated in response to stress that are adaptive. Persistent or dysregulated JAK/STAT3 signaling, however, is maladaptive and contributes to many diseases, including diseases characterized by chronic inflammation and fibrosis, and cancer. A comprehensive understanding of JAK/STAT3 signaling in normal development, and in adaptive and maladaptive responses to stress, is essential for the continued development of safe and effective therapies that target this signaling pathway., Competing Interests: Baylor College and Medicine, with D.J.T. as inventor, has filed 19 patents covering the use of TTI-101, a small-molecule inhibitor of STAT3 cited in this review. These patents are exclusively licensed to Tvardi Therapeutics, which was founded and is co-owned by D.J.T., (Copyright © 2020 by The Author(s).)
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- 2020
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20. Bempegaldesleukin selectively depletes intratumoral Tregs and potentiates T cell-mediated cancer therapy.
- Author
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Sharma M, Khong H, Fa'ak F, Bentebibel SE, Janssen LME, Chesson BC, Creasy CA, Forget MA, Kahn LMS, Pazdrak B, Karki B, Hailemichael Y, Singh M, Vianden C, Vennam S, Bharadwaj U, Tweardy DJ, Haymaker C, Bernatchez C, Huang S, Rajapakshe K, Coarfa C, Hurwitz ME, Sznol M, Hwu P, Hoch U, Addepalli M, Charych DH, Zalevsky J, Diab A, and Overwijk WW
- Subjects
- Animals, Antibodies, Monoclonal, Humanized administration & dosage, CD8-Positive T-Lymphocytes immunology, Carcinoma, Renal Cell genetics, Carcinoma, Renal Cell immunology, Cohort Studies, Drug Therapy, Combination, Female, Humans, Interferon-gamma genetics, Interferon-gamma immunology, Interleukin-2 administration & dosage, Interleukin-2 agonists, Interleukin-2 immunology, Ipilimumab administration & dosage, Lymphocyte Activation drug effects, Melanoma genetics, Melanoma immunology, Mice, Mice, Inbred C57BL, Receptors, Interleukin-2 genetics, Receptors, Interleukin-2 immunology, Tumor Necrosis Factor-alpha genetics, Tumor Necrosis Factor-alpha immunology, Carcinoma, Renal Cell drug therapy, Interleukin-2 analogs & derivatives, Melanoma drug therapy, Polyethylene Glycols administration & dosage, Prodrugs administration & dosage, T-Lymphocytes, Regulatory immunology
- Abstract
High dose interleukin-2 (IL-2) is active against metastatic melanoma and renal cell carcinoma, but treatment-associated toxicity and expansion of suppressive regulatory T cells (Tregs) limit its use in patients with cancer. Bempegaldesleukin (NKTR-214) is an engineered IL-2 cytokine prodrug that provides sustained activation of the IL-2 pathway with a bias to the IL-2 receptor CD122 (IL-2Rβ). Here we assess the therapeutic impact and mechanism of action of NKTR-214 in combination with anti-PD-1 and anti-CTLA-4 checkpoint blockade therapy or peptide-based vaccination in mice. NKTR-214 shows superior anti-tumor activity over native IL-2 and systemically expands anti-tumor CD8
+ T cells while inducing Treg depletion in tumor tissue but not in the periphery. Similar trends of intratumoral Treg dynamics are observed in a small cohort of patients treated with NKTR-214. Mechanistically, intratumoral Treg depletion is mediated by CD8+ Teff-associated cytokines IFN-γ and TNF-α. These findings demonstrate that NKTR-214 synergizes with T cell-mediated anti-cancer therapies.- Published
- 2020
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21. Combined Inhibition of STAT3 and DNA Repair in Palbociclib-Resistant ER-Positive Breast Cancer.
- Author
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Kettner NM, Vijayaraghavan S, Durak MG, Bui T, Kohansal M, Ha MJ, Liu B, Rao X, Wang J, Yi M, Carey JPW, Chen X, Eckols TK, Raghavendra AS, Ibrahim NK, Karuturi MS, Watowich SS, Sahin A, Tweardy DJ, Hunt KK, Tripathy D, and Keyomarsi K
- Subjects
- Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Antineoplastic Combined Chemotherapy Protocols adverse effects, Breast Neoplasms pathology, Cell Line, Tumor, Computational Biology methods, Epithelial-Mesenchymal Transition genetics, Female, Gene Expression Profiling, Humans, Immunohistochemistry, Piperazines pharmacology, Piperazines therapeutic use, Protein Kinase Inhibitors pharmacology, Pyridines pharmacology, Pyridines therapeutic use, Treatment Outcome, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Breast Neoplasms drug therapy, Breast Neoplasms metabolism, DNA Repair drug effects, Drug Resistance, Neoplasm, Receptors, Estrogen metabolism, STAT3 Transcription Factor antagonists & inhibitors
- Abstract
Purpose: Cyclin-dependent kinase 4/6 (CDK4/6) inhibitors are currently used in combination with endocrine therapy to treat advanced hormone receptor-positive, HER2-negative breast cancer. Although this treatment doubles time to progression compared with endocrine therapy alone, about 25%-35% of patients do not respond, and almost all patients eventually acquire resistance. Discerning the mechanisms of resistance to CDK4/6 inhibition is crucial in devising alternative treatment strategies., Experimental Design: Palbociclib-resistant cells (MCF-7 and T47D) were generated in a step-wise dose-escalading fashion. Whole-exome sequencing, genome-wide expression analysis, and proteomic analysis were performed in both resistant and parental (sensitive) cells. Pathway alteration was assessed mechanistically and pharmacologically. Biomarkers of altered pathways were examined in tumor samples from patients with palbociclib-treated breast cancer whose disease progressed while on treatment., Results: Palbociclib-resistant cells are cross-resistant to other CDK4/6 inhibitors and are also resistant to endocrine therapy (estrogen receptor downregulation). IL6/STAT3 pathway is induced, whereas DNA repair and estrogen receptor pathways are downregulated in the resistant cells. Combined inhibition of STAT3 and PARP significantly increased cell death in the resistant cells. Matched tumor samples from patients with breast cancer who progressed on palbociclib were examined for deregulation of estrogen receptor, DNA repair, and IL6/STAT3 signaling, and results revealed that these pathways are all altered as compared with the pretreatment tumor samples., Conclusions: Palbociclib resistance induces endocrine resistance, estrogen receptor downregulation, and alteration of IL6/STAT3 and DNA damage response pathways in cell lines and patient samples. Targeting IL6/STAT3 activity and DNA repair deficiency using a specific STAT3 inhibitor combined with a PARP inhibitor could effectively treat acquired resistance to palbociclib., (©2019 American Association for Cancer Research.)
- Published
- 2019
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22. Correction: Small-molecule inhibition of STAT3 in radioresistant head and neck squamous cell carcinoma.
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Bharadwaj U, Eckols TK, Xu X, Kasembeli MM, Chen Y, Adachi M, Song Y, Mo Q, Lai SY, and Tweardy DJ
- Abstract
[This corrects the article DOI: 10.18632/oncotarget.8368.].
- Published
- 2019
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23. Contribution of STAT3 to Inflammatory and Fibrotic Diseases and Prospects for its Targeting for Treatment.
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Kasembeli MM, Bharadwaj U, Robinson P, and Tweardy DJ
- Subjects
- Animals, Asthma immunology, Asthma metabolism, Cachexia immunology, Cachexia metabolism, Colitis, Ulcerative immunology, Crohn Disease immunology, Fibrosis immunology, Humans, Colitis, Ulcerative metabolism, Crohn Disease metabolism, Fibrosis metabolism, STAT3 Transcription Factor metabolism
- Abstract
Signal transducer and activator of transcription (STAT) 3 plays a central role in the host response to injury. It is activated rapidly within cells by many cytokines, most notably those in the IL-6 family, leading to pro-proliferative and pro-survival programs that assist the host in regaining homeostasis. With persistent activation, however, chronic inflammation and fibrosis ensue, leading to a number of debilitating diseases. This review summarizes advances in our understanding of the role of STAT3 and its targeting in diseases marked by chronic inflammation and/or fibrosis with a focus on those with the largest unmet medical need.
- Published
- 2018
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24. Intratumoral CD40 activation and checkpoint blockade induces T cell-mediated eradication of melanoma in the brain.
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Singh M, Vianden C, Cantwell MJ, Dai Z, Xiao Z, Sharma M, Khong H, Jaiswal AR, Faak F, Hailemichael Y, Janssen LME, Bharadwaj U, Curran MA, Diab A, Bassett RL, Tweardy DJ, Hwu P, and Overwijk WW
- Subjects
- Adenoviridae genetics, Animals, Brain pathology, CD4-CD8 Ratio, CD40 Antigens metabolism, CD40 Ligand genetics, CTLA-4 Antigen antagonists & inhibitors, Cell Line, Tumor, Enzyme Activation, Female, Immunotherapy methods, Mice, Mice, Inbred C57BL, Mice, Knockout, Programmed Cell Death 1 Receptor antagonists & inhibitors, Programmed Cell Death 1 Receptor biosynthesis, Brain Neoplasms immunology, Brain Neoplasms therapy, CD40 Antigens agonists, CD40 Ligand immunology, CD8-Positive T-Lymphocytes immunology, Melanoma, Experimental immunology, Melanoma, Experimental therapy
- Abstract
CD40 agonists bind the CD40 molecule on antigen-presenting cells and activate them to prime tumor-specific CD8
+ T cell responses. Here, we study the antitumor activity and mechanism of action of a nonreplicating adenovirus encoding a chimeric, membrane-bound CD40 ligand (ISF35). Intratumoral administration of ISF35 in subcutaneous B16 melanomas generates tumor-specific, CD8+ T cells that express PD-1 and suppress tumor growth. Combination therapy of ISF35 with systemic anti-PD-1 generates greater antitumor activity than each respective monotherapy. Triple combination of ISF35, anti-PD-1, and anti-CTLA-4 results in complete eradication of injected and noninjected subcutaneous tumors, as well as melanoma tumors in the brain. Therapeutic efficacy is associated with increases in the systemic level of tumor-specific CD8+ T cells, and an increased ratio of intratumoral CD8+ T cells to CD4+ Tregs. These results provide a proof of concept of systemic antitumor activity after intratumoral CD40 triggering with ISF35 in combination with checkpoint blockade for multifocal cancer, including the brain.- Published
- 2017
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25. Multifunctional Effects of a Small-Molecule STAT3 Inhibitor on NASH and Hepatocellular Carcinoma in Mice.
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Jung KH, Yoo W, Stevenson HL, Deshpande D, Shen H, Gagea M, Yoo SY, Wang J, Eckols TK, Bharadwaj U, Tweardy DJ, and Beretta L
- Subjects
- Animals, Antineoplastic Agents pharmacology, Biopsy, Carcinoma, Hepatocellular diagnosis, Carcinoma, Hepatocellular drug therapy, Cell Line, Tumor, Cell Proliferation drug effects, Cell Survival drug effects, Cytokines metabolism, Disease Models, Animal, Gene Expression Profiling, Humans, Immunohistochemistry, Inflammation Mediators, Liver Function Tests, Liver Neoplasms drug therapy, Liver Neoplasms pathology, Magnetic Resonance Imaging, Mice, Mice, Knockout, Non-alcoholic Fatty Liver Disease diagnosis, Non-alcoholic Fatty Liver Disease drug therapy, PTEN Phosphohydrolase genetics, PTEN Phosphohydrolase metabolism, Signal Transduction drug effects, Xenograft Model Antitumor Assays, Carcinoma, Hepatocellular metabolism, Liver Neoplasms metabolism, Naphthols pharmacology, Non-alcoholic Fatty Liver Disease metabolism, STAT3 Transcription Factor antagonists & inhibitors, Sulfonamides pharmacology
- Abstract
Purpose: The incidence of hepatocellular carcinoma is increasing in the United States, and liver cancer is the second leading cause of cancer-related mortality worldwide. Nonalcoholic steatohepatitis (NASH) is becoming an important risk for hepatocellular carcinoma, and most patients with hepatocellular carcinoma have underlying liver cirrhosis and compromised liver function, which limit treatment options. Thus, novel therapeutic strategies to prevent or treat hepatocellular carcinoma in the context of NASH and cirrhosis are urgently needed. Experimental Design: Constitutive activation of STAT3 is frequently detected in hepatocellular carcinoma tumors. STAT3 signaling plays a pivotal role in hepatocellular carcinoma survival, growth, angiogenesis, and metastasis. We identified C188-9, a novel small-molecule STAT3 inhibitor using computer-aided rational drug design. In this study, we evaluated the therapeutic potential of C188-9 for hepatocellular carcinoma treatment and prevention. Results: C188-9 showed antitumor activity in vitro in three hepatocellular carcinoma cell lines. In mice with hepatocyte-specific deletion of Pten (Hep Pten
- mice), C188-9 treatment blocked hepatocellular carcinoma tumor growth, reduced tumor development, and reduced liver steatosis, inflammation, and bile ductular reactions, resulting in improvement of the pathological lesions of NASH. Remarkably, C188-9 also greatly reduced liver injury in these mice as measured by serum aspartate aminotransferase and alanine transaminase levels. Analysis of gene expression showed that C188-9 treatment of Hep Pten- mice resulted in inhibition of signaling pathways downstream of STAT3, STAT1, TREM-1, and Toll-like receptors. In contrast, C188-9 treatment increased liver specification and differentiation gene pathways. Conclusions: Our results suggest that C188-9 should be evaluated further for the treatment and/or prevention of hepatocellular carcinoma. Clin Cancer Res; 23(18); 5537-46. ©2017 AACR ., (©2017 American Association for Cancer Research.)- Published
- 2017
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26. Protein stabilization improves STAT3 function in autosomal dominant hyper-IgE syndrome.
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Bocchini CE, Nahmod K, Katsonis P, Kim S, Kasembeli MM, Freeman A, Lichtarge O, Makedonas G, and Tweardy DJ
- Subjects
- Animals, CD4-Positive T-Lymphocytes drug effects, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes drug effects, CD8-Positive T-Lymphocytes immunology, Cytokines pharmacology, DNA-Binding Proteins metabolism, Diterpenes pharmacology, Half-Life, Heat Shock Transcription Factors, Herpesvirus 4, Human physiology, Humans, Interleukin-17 metabolism, Job Syndrome pathology, Mice, Models, Molecular, Mutant Proteins metabolism, Mutation genetics, Phosphotyrosine metabolism, Protein Binding drug effects, Protein Stability drug effects, STAT3 Transcription Factor chemistry, STAT3 Transcription Factor genetics, Spleen pathology, Transcription Factors metabolism, Job Syndrome metabolism, STAT3 Transcription Factor metabolism
- Abstract
Autosomal dominant hyper-IgE syndrome (AD-HIES) is caused by dominant-negative mutations in STAT3; however, the molecular basis for mutant STAT3 allele dysfunction is unclear and treatment remains supportive. We hypothesized that AD-HIES mutations decrease STAT3 protein stability and that mutant STAT3 activity can be improved by agents that increase chaperone protein activity. We used computer modeling to characterize the effect of STAT3 mutations on protein stability. We measured STAT3 protein half-life (t
1/2 ) and determined levels of STAT3 phosphorylated on tyrosine (Y) 705 (pY-STAT3) and mRNA levels of STAT3 gene targets in Epstein-Barr virus-transformed B (EBV) cells, human peripheral blood mononuclear cells (PBMCs), and mouse splenocytes incubated without or with chaperone protein modulators-HSF1A, a small-molecule TRiC modulator, or geranylgeranylacetone (GGA), a drug that upregulates heat shock protein (HSP) 70 and HSP90. Computer modeling predicted that 81% of AD-HIES mutations are destabilizing. STAT3 protein t1/2 in EBV cells from AD-HIES patients with destabilizing STAT3 mutations was markedly reduced. Treatment of EBV cells containing destabilizing STAT3 mutations with either HSF1A or GGA normalized STAT3 t1/2 , increased pY-STAT3 levels, and increased mRNA levels of STAT3 target genes up to 79% of control. In addition, treatment of human PBMCs or mouse splenocytes containing destabilizing STAT3 mutations with either HSF1A or GGA increased levels of cytokine-activated pY-STAT3 within human CD4+ and CD8+ T cells and numbers of IL-17-producing CD4+ mouse splenocytes, respectively. Thus, most AD-HIES STAT3 mutations are destabilizing; agents that modulate chaperone protein function improve STAT3 stability and activity in T cells and may provide a specific treatment.- Published
- 2016
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27. STAT3 inhibition prevents lung inflammation, remodeling, and accumulation of Th2 and Th17 cells in a murine asthma model.
- Author
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Gavino AC, Nahmod K, Bharadwaj U, Makedonas G, and Tweardy DJ
- Subjects
- Airway Remodeling, Allergens administration & dosage, Animals, Asthma pathology, Cytokines metabolism, Disease Models, Animal, Inflammation Mediators metabolism, Lung immunology, Lung metabolism, Lung pathology, Male, Mice, Pyroglyphidae immunology, T-Lymphocyte Subsets immunology, T-Lymphocyte Subsets metabolism, Asthma immunology, Asthma metabolism, STAT3 Transcription Factor antagonists & inhibitors, Th17 Cells immunology, Th17 Cells metabolism, Th2 Cells immunology, Th2 Cells metabolism
- Abstract
Background: STAT3 drives development of Th17 cells and cytokine production by Th2 and Th17 cells, which contribute to asthma. Alternative asthma treatments are needed, especially for the Th17 phenotype. We sought to determine whether C188-9, a small-molecule STAT3 inhibitor, can block Th2 and Th17 cell expansion and cytokine production to prevent house dust mite (HDM)-induced airway inflammation and remodeling., Methods: Three groups of C57BL/6 mice were treated intranasally (IN) and intraperitoneally (IP) daily for 3 weeks with the following: (i) vehicle 1 IN and vehicle 2 IP, (ii) HDM IN and vehicle 2 IP, or (iii) HDM IN and C188-9 IP. Sections of lung were stained with Alcian Blue/PAS and examined microscopically. Total (t) STAT3, STAT3 phosphorylated on Y705 (pSTAT3), IL-17, IL-13, IL-5, and IL-4 levels were measured in lung protein extracts and serum using Luminex beads. Frequencies of Th2-type and Th17-type lymphocytes were assessed in lungs and bronchoalveolar lavage fluid (BALF) by multiparametric flow cytometry., Results: HDM inhalation markedly increased airway goblet cell numbers and thickness of the epithelium and subepithelial smooth muscle layer, which was accompanied in the whole lung by increased pSTAT3, IL-4, IL-5, IL-13, and IL-17, and % CD4
+ T cells that produce IL-5, IL-13, and IL-17. HDM inhalation also increased serum IL-4 and IL-17 levels and increased BALF % CD4+ T cells that produce IL-5 and IL-13. Remarkably, treatment with C188-9 normalized each endpoint., Conclusion: HDM-induced airway inflammation, remodeling, and Th2/Th17-type cell accumulation involve STAT3 activation that can be prevented by C188-9 treatment., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)- Published
- 2016
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28. Diminution of signal transducer and activator of transcription 3 signaling inhibits vascular permeability and anaphylaxis.
- Author
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Hox V, O'Connell MP, Lyons JJ, Sackstein P, Dimaggio T, Jones N, Nelson C, Boehm M, Holland SM, Freeman AF, Tweardy DJ, Olivera A, Metcalfe DD, and Milner JD
- Subjects
- Adherens Junctions metabolism, Anaphylaxis diagnosis, Anaphylaxis genetics, Animals, Capillary Permeability drug effects, Capillary Permeability genetics, Cell Degranulation drug effects, Cell Degranulation immunology, Cytokines metabolism, Disease Models, Animal, Human Umbilical Vein Endothelial Cells, Humans, Immunoglobulin E immunology, Inflammation Mediators metabolism, Mast Cells metabolism, Mice, Mice, Knockout, Mutation, Receptors, Histamine immunology, Receptors, Histamine metabolism, STAT3 Transcription Factor antagonists & inhibitors, STAT3 Transcription Factor genetics, Skin Tests, beta Catenin metabolism, src-Family Kinases metabolism, Anaphylaxis immunology, Anaphylaxis metabolism, Capillary Permeability immunology, STAT3 Transcription Factor metabolism, Signal Transduction drug effects
- Abstract
Background: During IgE-mediated immediate hypersensitivity reactions, vascular endothelial cells permeabilize in response to mast cell mediators. We have demonstrated previously that patients and mice with signal transducer and activator of transcription 3 (STAT3) mutations (autosomal dominant hyper-IgE syndrome [AD-HIES]) are partially protected from anaphylaxis., Objectives: We sought to study the mechanism by which STAT3 contributes to anaphylaxis and determine whether small-molecule inhibition of STAT3 can prevent anaphylaxis., Methods: Using unaffected and STAT3-inhibited or genetic loss-of-function samples, we performed histamine skin prick tests, investigated the contribution of STAT3 to animal models of anaphylaxis, and measured endothelial cell permeability, gene and protein expression, and histamine receptor-mediated signaling., Results: Although mouse mast cell degranulation was minimally affected by STAT3 blockade, mast cell mediator-induced anaphylaxis was blunted in Stat3 mutant mice with AD-HIES and in wild-type mice subjected to small-molecule STAT3 inhibition. Histamine skin prick test responses were diminished in patients with AD-HIES. Human umbilical vein endothelial cells derived from patients with AD-HIES or treated with a STAT3 inhibitor did not signal properly through Src or cause appropriate dissolution of the adherens junctions made up of the proteins vascular endothelial-cadherin and β-catenin. Furthermore, we found that diminished STAT3 target microRNA17-92 expression in human umbilical vein endothelial cells from patients with AD-HIES is associated with increased phosphatase and tensin homolog (PTEN) expression, which inhibits Src, and increased E2F transcription factor 1 expression, which regulates β-catenin cellular dynamics., Conclusions: These data demonstrate that STAT3-dependent transcriptional activity regulates critical components for the architecture and functional dynamics of endothelial junctions, thus permitting vascular permeability., (Published by Elsevier Inc.)
- Published
- 2016
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29. Chaperonin TRiC/CCT Recognizes Fusion Oncoprotein AML1-ETO through Subunit-Specific Interactions.
- Author
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Roh SH, Kasembeli MM, Galaz-Montoya JG, Chiu W, and Tweardy DJ
- Subjects
- Blotting, Western, Chromatography, Gel, Cryoelectron Microscopy, DNA metabolism, Gold Compounds, HSP70 Heat-Shock Proteins metabolism, HeLa Cells, Humans, Imaging, Three-Dimensional, Mass Spectrometry, Metal Nanoparticles, Protein Domains, Protein Folding, Protein Multimerization, Protein Stability, RUNX1 Translocation Partner 1 Protein, Chaperonin Containing TCP-1 metabolism, Core Binding Factor Alpha 2 Subunit metabolism, Oncogene Proteins, Fusion metabolism
- Abstract
AML1-ETO is the translational product of a chimeric gene created by the stable chromosome translocation t (8;21)(q22;q22). It causes acute myeloid leukemia (AML) by dysregulating the expression of genes critical for myeloid cell development and differentiation and recently has been reported to bind multiple subunits of the mammalian cytosolic chaperonin TRiC (or CCT), primarily through its DNA binding domain (AML1-175). Through these interactions, TRiC plays an important role in the synthesis, folding, and activity of AML1-ETO. Using single-particle cryo-electron microscopy, we demonstrate here that a folding intermediate of AML1-ETO's DNA-binding domain (AML1-175) forms a stable complex with apo-TRiC. Our structure reveals that AML1-175 associates directly with a specific subset of TRiC subunits in the open conformation., (Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.)
- Published
- 2016
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30. Small-molecule inhibition of STAT3 in radioresistant head and neck squamous cell carcinoma.
- Author
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Bharadwaj U, Eckols TK, Xu X, Kasembeli MM, Chen Y, Adachi M, Song Y, Mo Q, Lai SY, and Tweardy DJ
- Subjects
- Animals, Antineoplastic Agents chemical synthesis, Cell Line, Tumor, Cell Proliferation drug effects, Drug Screening Assays, Antitumor, Humans, Mice, Mice, Nude, Naphthols chemical synthesis, Radiation Tolerance drug effects, Squamous Cell Carcinoma of Head and Neck, Sulfonamides chemical synthesis, Xenograft Model Antitumor Assays, Antineoplastic Agents pharmacology, Carcinoma, Squamous Cell drug therapy, Head and Neck Neoplasms drug therapy, Naphthols pharmacology, STAT3 Transcription Factor antagonists & inhibitors, Sulfonamides pharmacology
- Abstract
While STAT3 has been validated as a target for treatment of many cancers, including head and neck squamous cell carcinoma (HNSCC), a STAT3 inhibitor is yet to enter the clinic. We used the scaffold of C188, a small-molecule STAT3 inhibitor previously identified by us, in a hit-to-lead program to identify C188-9. C188-9 binds to STAT3 with high affinity and represents a substantial improvement over C188 in its ability to inhibit STAT3 binding to its pY-peptide ligand, to inhibit cytokine-stimulated pSTAT3, to reduce constitutive pSTAT3 activity in multiple HNSCC cell lines, and to inhibit anchorage dependent and independent growth of these cells. In addition, treatment of nude mice bearing xenografts of UM-SCC-17B, a radioresistant HNSCC line, with C188-9, but not C188, prevented tumor xenograft growth. C188-9 treatment modulated many STAT3-regulated genes involved in oncogenesis and radioresistance, as well as radioresistance genes regulated by STAT1, due to its potent activity against STAT1, in addition to STAT3. C188-9 was well tolerated in mice, showed good oral bioavailability, and was concentrated in tumors. Thus, C188-9, either alone or in combination with radiotherapy, has potential for use in treating HNSCC tumors that demonstrate increased STAT3 and/or STAT1 activation., Competing Interests: Please note that DJT has the following conflict of interest: Baylor College of Medicine, with DJT as primary inventor, filed composition and methods of use patents on C188 and C188-9. StemMed, Ltd. currently holds an exclusive license to these compounds. DJT is founding partner, President, and CEO of StemMed and also have majority ownership of StemMed. He has been in compliance with all conflict of interests (COI) policies at Baylor College of Medicine and currently is in compliance with all COI policies at StemMed and at the University of Texas M.D. Anderson Cancer Center, where he relocated on December 1, 2014.
- Published
- 2016
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31. PARP Inhibition Suppresses Growth of EGFR-Mutant Cancers by Targeting Nuclear PKM2.
- Author
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Li N, Feng L, Liu H, Wang J, Kasembeli M, Tran MK, Tweardy DJ, Lin SH, and Chen J
- Abstract
Upon growth factor stimulation or in some EGFR mutant cancer cells, PKM2 translocates into the nucleus to induce glycolysis and cell growth. Here, we report that nuclear PKM2 binds directly to poly-ADP ribose, and this PAR-binding capability is critical for its nuclear localization. Accordingly, PARP inhibition prevents nuclear retention of PKM2 and therefore suppresses cell proliferation and tumor growth. In addition, we found that PAR level correlates with nuclear localization of PKM2 in EGFR mutant brain and lung cancers, suggesting that PAR-dependent nuclear localization of PKM2 likely contributes to tumor progression in EGFR mutant glioblastoma and lung cancers. In addition, some EGFR-inhibitor-resistant lung cancer cells are sensitive to PARP inhibitors. Taken together, our data indicate that suppression of PKM2 nuclear function by PARP inhibitors represents a treatment strategy for EGFR-inhibitor-resistant cancers., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2016
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32. Association Between Hepatitis C Virus and Head and Neck Cancers.
- Author
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Mahale P, Sturgis EM, Tweardy DJ, Ariza-Heredia EJ, and Torres HA
- Subjects
- Aged, Case-Control Studies, Esophageal Neoplasms etiology, Female, Humans, Hypopharyngeal Neoplasms blood, Laryngeal Neoplasms blood, Lung Neoplasms etiology, Male, Middle Aged, Mouth Neoplasms blood, Nasopharyngeal Neoplasms blood, Oropharyngeal Neoplasms blood, Papillomaviridae immunology, Seroepidemiologic Studies, Smoking adverse effects, Urinary Bladder Neoplasms etiology, Esophageal Neoplasms blood, Hepacivirus immunology, Hepatitis C Antibodies blood, Hypopharyngeal Neoplasms epidemiology, Laryngeal Neoplasms epidemiology, Lung Neoplasms blood, Mouth Neoplasms epidemiology, Nasopharyngeal Neoplasms epidemiology, Oropharyngeal Neoplasms epidemiology, Urinary Bladder Neoplasms blood
- Abstract
Background: Hepatitis C virus (HCV) infection is associated with hepatocellular carcinoma and non-Hodgkin's lymphoma. In 2009, MD Anderson established the first US clinic for treating HCV-infected cancer patients, where we observed an unexpectedly large number of patients with head and neck cancers (HNCs). We sought to determine whether HCV is associated with HNCs., Methods: In this case-control study, medical records of cancer patients tested for HCV antibodies at our center from 2004 through 2014 were identified. Case subjects had new-onset primary oropharyngeal or nonoropharyngeal (oral cavity, nasopharynx, hypopharynx, or larynx) HNCs. Control subjects had smoking-associated (lung, esophagus, or urinary bladder) cancers. Biopsy reports of oropharyngeal cancers tested for human papillomavirus (HPV) were reviewed. Patients with lymphoma were excluded. Multivariable logistic regression models were constructed. All statistical tests were two-sided., Results: Of 34 545 cancer patients tested for HCV antibodies, 409 case subjects (164 oropharyngeal and 245 nonoropharyngeal) and 694 control subjects (378 lung, 168 esophagus, and 148 urinary bladder) were studied. The prevalence of HCV seropositivity was higher in oropharyngeal cancer patients (14.0%, 95% confidence interval [CI] = 8.7% to 19.4%, vs 6.5%, 95% CI = 4.6% to 8.3%), particularly HPV-positive oropharyngeal cancer patients (16.9%, 95% CI = 8.7% to 24.9%, vs 6.5%, 95% CI = 4.6% to 8.3%), and nonoropharyngeal HNC patients (20.0%, 95% CI = 14.9% to 25.0%, vs 6.5%, 95% CI = 4.6% to 8.3%) than in control subjects. Adjusted models showed a statistically significant association of HCV seropositivity with nonoropharyngeal (except nasopharyngeal) HNCs (odds ratio [OR] = 2.85, 95% CI = 1.38 to 5.88) and HPV-positive oropharyngeal cancers (OR = 2.97, 95% CI = 1.31 to 6.76)., Conclusions: HCV is associated with nonoropharyngeal (except nasopharyngeal) and HPV-positive oropharyngeal HNCs. Further studies are required to explore the possible interaction between HCV and HPV, and the association between HCV and other HPV-related malignancies., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2016
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33. Chaperonin TRiC/CCT Modulates the Folding and Activity of Leukemogenic Fusion Oncoprotein AML1-ETO.
- Author
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Roh SH, Kasembeli M, Galaz-Montoya JG, Trnka M, Lau WC, Burlingame A, Chiu W, and Tweardy DJ
- Subjects
- Animals, Cattle, Cell Survival, Chaperonin Containing TCP-1 antagonists & inhibitors, Chaperonin Containing TCP-1 chemistry, Core Binding Factor Alpha 2 Subunit chemistry, Core Binding Factor Alpha 2 Subunit genetics, HEK293 Cells, Humans, Immunoprecipitation, Neoplasm Proteins antagonists & inhibitors, Neoplasm Proteins chemistry, Oncogene Proteins, Fusion chemistry, Oncogene Proteins, Fusion genetics, Peptide Fragments chemistry, Peptide Fragments genetics, Peptide Fragments metabolism, Protein Conformation, Protein Folding, Protein Interaction Domains and Motifs, Protein Stability, Protein Subunits, RUNX1 Translocation Partner 1 Protein, Rats, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins metabolism, Reticulocytes metabolism, Chaperonin Containing TCP-1 metabolism, Core Binding Factor Alpha 2 Subunit metabolism, Gene Expression Regulation, Neoplastic, HSP70 Heat-Shock Proteins metabolism, Models, Molecular, Neoplasm Proteins metabolism, Oncogene Proteins, Fusion metabolism
- Abstract
AML1-ETO is the most common fusion oncoprotein causing acute myeloid leukemia (AML), a disease with a 5-year survival rate of only 24%. AML1-ETO functions as a rogue transcription factor, altering the expression of genes critical for myeloid cell development and differentiation. Currently, there are no specific therapies for AML1-ETO-positive AML. While known for decades to be the translational product of a chimeric gene created by the stable chromosome translocation t(8;21)(q22;q22), it is not known how AML1-ETO achieves its native and functional conformation or whether this process can be targeted for therapeutic benefit. Here, we show that the biosynthesis and folding of the AML1-ETO protein is facilitated by interaction with the essential eukaryotic chaperonin TRiC (or CCT). We demonstrate that a folding intermediate of AML1-ETO binds to TRiC directly, mainly through its β-strand rich, DNA-binding domain (AML-(1-175)), with the assistance of HSP70. Our results suggest that TRiC contributes to AML1-ETO proteostasis through specific interactions between the oncoprotein's DNA-binding domain, which may be targeted for therapeutic benefit., (© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2016
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34. The Opposing Function of STAT3 as an Oncoprotein and Tumor Suppressor Is Dictated by the Expression Status of STAT3β in Esophageal Squamous Cell Carcinoma.
- Author
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Zhang HF, Chen Y, Wu C, Wu ZY, Tweardy DJ, Alshareef A, Liao LD, Xue YJ, Wu JY, Chen B, Xu XE, Gopal K, Gupta N, Li EM, Xu LY, and Lai R
- Subjects
- Animals, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Carcinoma, Squamous Cell diagnosis, Carcinoma, Squamous Cell drug therapy, Carcinoma, Squamous Cell mortality, Cell Line, Tumor, Disease Models, Animal, Drug Resistance, Neoplasm genetics, Esophageal Neoplasms diagnosis, Esophageal Neoplasms drug therapy, Esophageal Neoplasms mortality, Esophageal Squamous Cell Carcinoma, Humans, Immunohistochemistry, Models, Biological, Neoplastic Stem Cells metabolism, Phosphorylation, Prognosis, Protein Multimerization, Protein Tyrosine Phosphatases metabolism, STAT3 Transcription Factor chemistry, Transcriptional Activation, Xenograft Model Antitumor Assays, Carcinoma, Squamous Cell genetics, Carcinoma, Squamous Cell metabolism, Esophageal Neoplasms genetics, Esophageal Neoplasms metabolism, Gene Expression Regulation, Neoplastic, Oncogene Proteins, STAT3 Transcription Factor genetics, STAT3 Transcription Factor metabolism, Tumor Suppressor Proteins
- Abstract
Purpose: STAT3 is known to have both oncogenic and tumor suppressive effects, but the regulation of these opposing effects is elusive. We hypothesized that STAT3β, one of the two STAT3 isoforms, is the key determinant in this context., Experimental Design: The prognostic significance of STAT3β and phospho-STAT3α(Y705) (pSTAT3α(Y705)) was evaluated in 286 cases of patients with esophageal squamous cell carcinoma (ESCC). STAT3β-induced changes in the chemosensitivity to cisplatin and 5-fluorouracil were assessed both in vitro and in vivo. STAT3β-induced changes in the frequency of cancer stem cells were evaluated using Hoechst and CD44 staining. How STAT3β regulates STAT3α was determined using immunoprecipitation, confocal microscopy, DNA-binding, and chromatin immunoprecipitation-PCR., Results: STAT3β expression is an independent protective prognostic marker in patients with ESCC, which strongly correlated with longer overall survival (P = 0.0009) and recurrence-free survival (P = 0.0001). STAT3β significantly decreased the cancer stem cell population, and sensitized ESCC cells to cisplatin and 5-fluorouracil in tumor xenografts. Mechanistically, STAT3β markedly attenuated the transcription activity of STAT3α via inducing STAT3α:STAT3β heterodimers. However, the heterodimer formation decreased the binding between STAT3α and PTPN9 (better known as PTP-MEG2), a protein tyrosine phosphatase, thereby promoting the phosphorylation of STAT3α(Y705) and enhancing its nuclear translocation and DNA binding. Correlating with this, high STAT3β expression converts the prognostic value of pSTAT3α(Y705) from unfavorable to favorable in patients with ESCC., Conclusions: STAT3β suppresses chemoresistance and cancer stemness by blocking the transcriptional activity of STAT3α. The paradoxical increase in pSTAT3α(Y705) induced by STAT3β carries important implications as to how the biologic and prognostic significance of STAT3 in cancers should be interpreted., (©2015 American Association for Cancer Research.)
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- 2016
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35. Correction: Piperlongumine Blocks JAK2-STAT3 to Inhibit Collagen-Induced Platelet Reactivity Independent of Reactive Oxygen Species†.
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Yuan H, Houck KL, Tian Y, Bharadwaj U, Hull K, Zhou Z, Zhu M, Wu X, Tweardy DJ, Romo D, Fu X, Zhang Y, Zhang J, and Dong JF
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- 2016
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36. A mix of S and ΔS variants of STAT3 enable survival of activated B-cell-like diffuse large B-cell lymphoma cells in culture.
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Zheng M, Turton KB, Zhu F, Li Y, Grindle KM, Annis DS, Lu L, Drennan AC, Tweardy DJ, Bharadwaj U, Mosher DF, and Rui L
- Abstract
Activated B-cell-like diffuse large B-cell lymphoma (ABC DLBCL) is characterized by increased expression and activator of signal transducer and activator of transcription 3 (STAT3). ABC DLBCL cells require STAT3 for growth in culture. In ABC DLBCL cells, eosinophils and perhaps all cells, four variant STAT3 mRNAs (Sα, ΔSα, Sβ and ΔSβ) are present as a result of two alternative splicing events, one that results in the inclusion of a 55-residue C-terminal transactivation domain (α) or a truncated C-terminal domain with 7 unique residues (β) and a second that includes (S) or excludes (ΔS) the codon for Ser-701 in the linker between the SH2 and C-terminal domains. A substantial literature indicates that both α and β variants are required for optimal STAT3 function, but nothing is known about functions of ΔS variants. We used a knockdown/re-expression strategy to explore whether survival of ABC DLBCL cells requires that the four variants be in an appropriate ratio. No single variant rescued survival as well as STAT3Sα-C, Sα with activating mutations (A661C and N663C) in the SH2 domain. Better rescue was achieved when all four variants were re-expressed or Sα and ΔSα or Sβ and ΔSβ were re-expressed in pairs. Rescue correlated with expression of STAT3-sensitive genes NFKBIA and NFKBIZ. We consider a variety of explanations why a mix of S and ΔS variants of STAT3 should enable survival of ABC DLBCL cells.
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- 2016
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37. Substance P Receptor Signaling Mediates Doxorubicin-Induced Cardiomyocyte Apoptosis and Triple-Negative Breast Cancer Chemoresistance.
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Robinson P, Kasembeli M, Bharadwaj U, Engineer N, Eckols KT, and Tweardy DJ
- Subjects
- Animals, Apoptosis drug effects, Aprepitant, Cardiotoxicity genetics, Cardiotoxicity pathology, Cell Line, Tumor, Doxorubicin administration & dosage, Drug Resistance, Neoplasm genetics, Humans, Morpholines administration & dosage, Myocytes, Cardiac drug effects, Myocytes, Cardiac metabolism, Rats, Triple Negative Breast Neoplasms drug therapy, Triple Negative Breast Neoplasms genetics, Triple Negative Breast Neoplasms pathology, Cardiotoxicity drug therapy, Doxorubicin adverse effects, Receptors, Neurokinin-1 genetics, Triple Negative Breast Neoplasms complications
- Abstract
Doxorubicin (DOX), an anthracycline, is broadly considered the most active single agent available for treating breast cancer but has been known to induce cardiotoxicity. Although DOX is highly effective in treating triple-negative breast cancer (TNBC), DOX can have poor outcomes owing to induction of chemoresistance. There is an urgent need to develop new therapies for TNBC aimed at improving DOX outcome and DOX-induced cardiotoxicity. Substance P (SP), a neuropeptide involved in pain transmission is known to stimulate production of reactive oxygen species (ROS). Elevated cardiac ROS is linked with heart injury and failure. We investigated the role of SP in chemotherapy-associated death of cardiomyocytes and chemoresistance. We showed that pretreating a cardiomyocyte cell line (H9C2) and a TNBC cell line (MDA-MB 231) with aprepitant, a SP receptor antagonist that is routinely used to treat chemotherapy-associated associated nausea, decreased DOX-induced reduction of cell viability, apoptotic cell death, and ROS production in cardiomyocytes and increased DOX-induced reduction of cell viability, apoptotic cell death, and ROS production in TNBC cells compared with cells treated with DOX alone. Our findings demonstrate the ability of aprepitant to decrease DOX-induced killing of cardiomyocytes and to increase cancer cell sensitivity to DOX, which has tremendous clinical significance.
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- 2016
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38. STAT-3 contributes to pulmonary fibrosis through epithelial injury and fibroblast-myofibroblast differentiation.
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Pedroza M, Le TT, Lewis K, Karmouty-Quintana H, To S, George AT, Blackburn MR, Tweardy DJ, and Agarwal SK
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- Animals, Bleomycin pharmacology, Cell Differentiation drug effects, Disease Models, Animal, Idiopathic Pulmonary Fibrosis genetics, Male, Mice, Inbred C57BL, STAT3 Transcription Factor genetics, Cell Differentiation physiology, Epithelial Cells metabolism, Idiopathic Pulmonary Fibrosis metabolism, Lung metabolism, Myofibroblasts metabolism, STAT3 Transcription Factor metabolism
- Abstract
Lung fibrosis is the hallmark of the interstitial lung diseases. Alveolar epithelial cell (AEC) injury is a key step that contributes to a profibrotic microenvironment. Fibroblasts and myofibroblasts subsequently accumulate and deposit excessive extracellular matrix. In addition to TGF-β, the IL-6 family of cytokines, which signal through STAT-3, may also contribute to lung fibrosis. In the current manuscript, the extent to which STAT-3 inhibition decreases lung fibrosis is investigated. Phosphorylated STAT-3 was elevated in lung biopsies from patients with idiopathic pulmonary fibrosis and bleomycin (BLM)-induced fibrotic murine lungs. C-188-9, a small molecule STAT-3 inhibitor, decreased pulmonary fibrosis in the intraperitoneal BLM model as assessed by arterial oxygen saturation (control, 84.4 ± 1.3%; C-188-9, 94.4 ± 0.8%), histology (Ashcroft score: untreated, 5.4 ± 0.25; C-188-9, 3.3 ± 0.14), and attenuated fibrotic markers such as diminished α-smooth muscle actin, reduced collagen deposition. In addition, C-188-9 decreased the expression of epithelial injury markers, including hypoxia-inducible factor-1α (HIF-1α) and plasminogen activator inhibitor-1 (PAI-1). In vitro studies show that inhibition of STAT-3 decreased IL-6- and TGF-β-induced expression of multiple genes, including HIF-1α and PAI-1, in AECs. Furthermore, C-188-9 decreased fibroblast-to-myofibroblast differentiation. Finally, TGF-β stimulation of lung fibroblasts resulted in SMAD2/SMAD3-dependent phosphorylation of STAT-3. These findings demonstrate that STAT-3 contributes to the development of lung fibrosis and suggest that STAT-3 may be a therapeutic target in pulmonary fibrosis., (© FASEB.)
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- 2016
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39. Piperlongumine Blocks JAK2-STAT3 to Inhibit Collagen-Induced Platelet Reactivity Independent of Reactive Oxygen Species.
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Yuan H, Houck KL, Tian Y, Bharadwaj U, Hull K, Zhou Z, Zhu M, Wu X, Tweardy DJ, Romo D, Fu X, Zhang Y, Zhang J, and Dong JF
- Subjects
- Humans, Collagen antagonists & inhibitors, Dioxolanes pharmacology, Janus Kinase 2 antagonists & inhibitors, Platelet Activation drug effects, Platelet Aggregation drug effects, Reactive Oxygen Species metabolism, STAT3 Transcription Factor antagonists & inhibitors
- Abstract
Background: Piperlongumine (PL) is a compound isolated from the piper longum plant. It possesses anti-cancer activities through blocking the transcription factor STAT3 and by inducing reactive oxygen species (ROS) in cancer, but not normal cells. It also inhibits platelet aggregation induced by collagen, but the underlying mechanism is not known., Objective: We conducted in vitro experiments to test the hypothesis that PL regulates a non-transcriptional activity of STAT3 to specifically reduce the reactivity of human platelets to collagen., Results: PL dose-dependently blocked collagen-induced platelet aggregation, calcium influx, CD62p expression and thrombus formation on collagen with a maximal inhibition at 100 μM. It reduced platelet microvesiculation induced by collagen. PL blocked the activation of JAK2 and STAT3 in collagen-stimulated platelets. This inhibitory effect was significantly reduced in platelets pretreated with a STAT3 inhibitor. Although PL induced ROS production in platelets; quenching ROS using excessive reducing agents: 20 μM GSH and 0.5 mM L-Cysteine, did not block the inhibitory effects. The NADPH oxidase inhibitor Apocynin also had no effect., Conclusions: PL inhibited collagen-induced platelet reactivity by targeting the JAK2-STAT3 pathway. We also provide experimental evidence that PL and collagen induce different oxidants that have differential effects on platelets. Studying these differential effects may uncover new mechanisms of regulating platelet functions by oxidants in redox signals.
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- 2015
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40. Contribution of the Type II Chaperonin, TRiC/CCT, to Oncogenesis.
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Roh SH, Kasembeli M, Bakthavatsalam D, Chiu W, and Tweardy DJ
- Subjects
- Apoptosis, Carcinogenesis metabolism, Carcinogenesis pathology, Chaperonin Containing TCP-1 metabolism, Disease Progression, Genomic Instability, HSP70 Heat-Shock Proteins genetics, HSP70 Heat-Shock Proteins metabolism, HSP90 Heat-Shock Proteins genetics, HSP90 Heat-Shock Proteins metabolism, Humans, Neoplasm Proteins genetics, Neoplasm Proteins metabolism, Neoplasms metabolism, Neoplasms pathology, Protein Conformation, Protein Folding, Signal Transduction, Carcinogenesis genetics, Chaperonin Containing TCP-1 genetics, Gene Expression Regulation, Neoplastic, Neoplasm Proteins chemistry, Neoplasms genetics
- Abstract
The folding of newly synthesized proteins and the maintenance of pre-existing proteins are essential in sustaining a living cell. A network of molecular chaperones tightly guides the folding, intracellular localization, and proteolytic turnover of proteins. Many of the key regulators of cell growth and differentiation have been identified as clients of molecular chaperones, which implies that chaperones are potential mediators of oncogenesis. In this review, we briefly provide an overview of the role of chaperones, including HSP70 and HSP90, in cancer. We further summarize and highlight the emerging the role of chaperonin TRiC (T-complex protein-1 ring complex, also known as CCT) in the development and progression of cancer mediated through its critical interactions with oncogenic clients that modulate growth deregulation, apoptosis, and genome instability in cancer cells. Elucidation of how TRiC modulates the folding and function of oncogenic clients will provide strategies for developing novel cancer therapies.
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- 2015
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41. Small-molecule targeting of signal transducer and activator of transcription (STAT) 3 to treat non-small cell lung cancer.
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Lewis KM, Bharadwaj U, Eckols TK, Kolosov M, Kasembeli MM, Fridley C, Siller R, and Tweardy DJ
- Subjects
- Animals, Apoptosis drug effects, Apoptosis genetics, Carcinoma, Non-Small-Cell Lung genetics, Cell Line, Cell Line, Tumor, Cell Survival drug effects, Cell Survival genetics, Gene Expression Regulation, Neoplastic drug effects, Gene Expression Regulation, Neoplastic genetics, Humans, Lung drug effects, Lung Neoplasms genetics, Mice, Mice, Nude, Phosphorylation drug effects, Phosphorylation genetics, RNA, Small Interfering genetics, Signal Transduction genetics, Transcriptional Activation genetics, Antineoplastic Agents pharmacology, Carcinoma, Non-Small-Cell Lung drug therapy, Lung Neoplasms drug therapy, STAT3 Transcription Factor genetics, Signal Transduction drug effects, Transcriptional Activation drug effects
- Abstract
Objective: Lung cancer is the leading cause of cancer death in both men and women. Non-small cell lung cancer (NSCLC) has an overall 5-year survival rate of 15%. While aberrant STAT3 activation has previously been observed in NSCLC, the scope of its contribution is uncertain and agents that target STAT3 for treatment are not available clinically., Methods: We determined levels of activated STAT3 (STAT3 phosphorylated on Y705, pSTAT3) and the two major isoforms of STAT3 (α and β) in protein extracts of 8 NSCLC cell lines, as well as the effects of targeting STAT3 in vitro and in vivo in NSCLC cells using short hairpin (sh) RNA and two novel small-molecule STAT3 inhibitors, C188-9 and piperlongumine (PL)., Results: Levels of pSTAT3, STAT3α, and STATβ were increased in 7 of 8 NSCLC cell lines. Of note, levels of pSTAT3 were tightly correlated with levels of STAT3β, but not STAT3α. Targeting of STAT3 in A549 cells using shRNA decreased tSTAT3 by 75%; this was accompanied by a 47-78% reduction in anchorage-dependent and anchorage-independent growth and a 28-45% reduction in mRNA levels for anti-apoptotic STAT3 gene targets. C188-9 and PL (@30 μM) each reduced pSTAT3 levels in all NSCLC cell lines tested by ≥50%, reduced anti-apoptotic protein mRNA levels by 25-60%, and reduced both anchorage-dependent and anchorage-independent growth of NSCLC cell lines with IC50 values ranging from 3.06 to 52.44 μM and 0.86 to 11.66 μM, respectively. Treatment of nude mice bearing A549 tumor xenografts with C188-9 or PL blocked tumor growth and reduced levels of pSTAT3 and mRNA encoding anti-apoptotic proteins., Conclusion: STAT3 is essential for growth of NSCLC cell lines and tumors and its targeting using C188-9 or PL may be a useful strategy for treatment., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)
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- 2015
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42. Rhodium(II) Proximity-Labeling Identifies a Novel Target Site on STAT3 for Inhibitors with Potent Anti-Leukemia Activity.
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Minus MB, Liu W, Vohidov F, Kasembeli MM, Long X, Krueger MJ, Stevens A, Kolosov MI, Tweardy DJ, Sison EA, Redell MS, and Ball ZT
- Subjects
- Animals, Antineoplastic Agents chemistry, Apoptosis drug effects, Binding Sites drug effects, Catalysis, Cell Proliferation drug effects, Dose-Response Relationship, Drug, Drug Screening Assays, Antitumor, Humans, Leukemia, Myeloid, Acute pathology, Mice, Naphthalenes chemistry, Neoplasms, Experimental drug therapy, Neoplasms, Experimental pathology, STAT3 Transcription Factor metabolism, Structure-Activity Relationship, Sulfonamides chemistry, Antineoplastic Agents pharmacology, Leukemia, Myeloid, Acute drug therapy, Naphthalenes pharmacology, Rhodium chemistry, STAT3 Transcription Factor antagonists & inhibitors, Sulfonamides pharmacology
- Abstract
Nearly 40 % of children with acute myeloid leukemia (AML) suffer relapse arising from chemoresistance, often involving upregulation of the oncoprotein STAT3 (signal transducer and activator of transcription 3). Herein, rhodium(II)-catalyzed, proximity-driven modification identifies the STAT3 coiled-coil domain (CCD) as a novel ligand-binding site, and we describe a new naphthalene sulfonamide inhibitor that targets the CCD, blocks STAT3 function, and halts its disease-promoting effects in vitro, in tumor growth models, and in a leukemia mouse model, validating this new therapeutic target for resistant AML., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2015
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43. Inhibition of Stat3 activation suppresses caspase-3 and the ubiquitin-proteasome system, leading to preservation of muscle mass in cancer cachexia.
- Author
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Silva KA, Dong J, Dong Y, Dong Y, Schor N, Tweardy DJ, Zhang L, and Mitch WE
- Subjects
- Animals, CCAAT-Enhancer-Binding Protein-delta genetics, CCAAT-Enhancer-Binding Protein-delta metabolism, Cachexia genetics, Cachexia pathology, Carcinoma, Lewis Lung genetics, Carcinoma, Lewis Lung pathology, Caspase 3 genetics, Cell Line, Tumor, Colonic Neoplasms genetics, Colonic Neoplasms pathology, Mice, Mice, Knockout, Muscle Proteins genetics, Muscle Proteins metabolism, Muscle, Skeletal pathology, Myostatin genetics, Myostatin metabolism, Proteasome Endopeptidase Complex genetics, Proteolysis, SKP Cullin F-Box Protein Ligases genetics, SKP Cullin F-Box Protein Ligases metabolism, STAT3 Transcription Factor genetics, Tripartite Motif Proteins, Ubiquitin genetics, Ubiquitin-Protein Ligases genetics, Ubiquitin-Protein Ligases metabolism, Cachexia metabolism, Carcinoma, Lewis Lung metabolism, Caspase 3 metabolism, Colonic Neoplasms metabolism, Muscle, Skeletal metabolism, Proteasome Endopeptidase Complex metabolism, STAT3 Transcription Factor metabolism, Ubiquitin metabolism
- Abstract
Cachexia occurs in patients with advanced cancers. Despite the adverse clinical impact of cancer-induced muscle wasting, pathways causing cachexia are controversial, and clinically reliable therapies are not available. A trigger of muscle protein loss is the Jak/Stat pathway, and indeed, we found that conditioned medium from C26 colon carcinoma (C26) or Lewis lung carcinoma cells activates Stat3 (p-Stat3) in C2C12 myotubes. We identified two proteolytic pathways that are activated in muscle by p-Stat3; one is activation of caspase-3, and the other is p-Stat3 to myostatin, MAFbx/Atrogin-1, and MuRF-1 via CAAT/enhancer-binding protein δ (C/EBPδ). Using sequential deletions of the caspase-3 promoter and CHIP assays, we determined that Stat3 activation increases caspase-3 expression in C2C12 cells. Caspase-3 expression and proteolytic activity were stimulated by p-Stat3 in muscles of tumor-bearing mice. In mice with cachexia caused by Lewis lung carcinoma or C26 tumors, knock-out of p-Stat3 in muscle or with a small chemical inhibitor of p-Stat3 suppressed muscle mass losses, improved protein synthesis and degradation in muscle, and increased body weight and grip strength. Activation of p-Stat3 stimulates a pathway from C/EBPδ to myostatin and expression of MAFbx/Atrogin-1 and increases the ubiquitin-proteasome system. Indeed, C/EBPδ KO decreases the expression of MAFbx/Atrogin-1 and myostatin, while increasing muscle mass and grip strength. In conclusion, cancer stimulates p-Stat3 in muscle, activating protein loss by stimulating caspase-3, myostatin, and the ubiquitin-proteasome system. These results could lead to novel strategies for preventing cancer-induced muscle wasting., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)
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- 2015
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44. Drug-repositioning screening identified piperlongumine as a direct STAT3 inhibitor with potent activity against breast cancer.
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Bharadwaj U, Eckols TK, Kolosov M, Kasembeli MM, Adam A, Torres D, Zhang X, Dobrolecki LE, Wei W, Lewis MT, Dave B, Chang JC, Landis MD, Creighton CJ, Mancini MA, and Tweardy DJ
- Subjects
- Active Transport, Cell Nucleus drug effects, Animals, Antineoplastic Agents adverse effects, Antineoplastic Agents therapeutic use, Apoptosis drug effects, Cell Proliferation drug effects, Dioxolanes adverse effects, Dioxolanes therapeutic use, Female, Gene Expression drug effects, Humans, MCF-7 Cells, Mice, Mice, Nude, Mice, SCID, Neoplasm Transplantation, Oxidative Stress drug effects, Phosphorylation drug effects, Reverse Transcriptase Polymerase Chain Reaction, STAT3 Transcription Factor genetics, STAT3 Transcription Factor metabolism, Signal Transduction drug effects, Spheroids, Cellular drug effects, Surface Plasmon Resonance, Transplantation, Heterologous, Tumor Cells, Cultured, Antineoplastic Agents pharmacology, Breast Neoplasms drug therapy, Dioxolanes pharmacology, Drug Repositioning, STAT3 Transcription Factor antagonists & inhibitors
- Abstract
Signal transducer and activator of transcription (STAT) 3 regulates many cardinal features of cancer including cancer cell growth, apoptosis resistance, DNA damage response, metastasis, immune escape, tumor angiogenesis, the Warburg effect and oncogene addiction and has been validated as a drug target for cancer therapy. Several strategies have been used to identify agents that target Stat3 in breast cancer but none has yet entered into clinical use. We used a high-throughput fluorescence microscopy search strategy to identify compounds in a drug-repositioning library (Prestwick library) that block ligand-induced nuclear translocation of Stat3 and identified piperlongumine (PL), a natural product isolated from the fruit of the pepper Piper longum. PL inhibited Stat3 nuclear translocation, inhibited ligand-induced and constitutive Stat3 phosphorylation, and modulated expression of multiple Stat3-regulated genes. Surface plasmon resonance assay revealed that PL directly inhibited binding of Stat3 to its phosphotyrosyl peptide ligand. Phosphoprotein antibody array analysis revealed that PL does not modulate kinases known to activate Stat3 such as Janus kinases, Src kinase family members or receptor tyrosine kinases. PL inhibited anchorage-independent and anchorage-dependent growth of multiple breast cancer cell lines having increased pStat3 or total Stat3, and induced apoptosis. PL also inhibited mammosphere formation by tumor cells from patient-derived xenografts. PL's antitumorigenic function was causally linked to its Stat3-inhibitory effect. PL was non-toxic in mice up to a dose of 30 mg/kg/day for 14 days and caused regression of breast cancer cell line xenografts in nude mice. Thus, PL represents a promising new agent for rapid entry into the clinic for use in treating breast cancer, as well as other cancers in which Stat3 has a role.
- Published
- 2015
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45. First Description of Oral Cryptococcus neoformans Causing Osteomyelitis of the Mandible, Manubrium and Third Rib with Associated Soft Tissue Abscesses in an Immunocompetent Host.
- Author
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DiNardo AR, Schmidt D, Mitchell A, Kaufman Y, and Tweardy DJ
- Abstract
The majority of disseminated cryptococcal infections occur in patients with acquired immunodeficiency syndrome (AIDS), with only 11-14% of cases occurring in patients without AIDS. Most non-AIDS related cases (75%) occur in patients with another immune deficiency. Here, we present the first case of mucocutaneous cryptococcal disease in an immunocompetent host, review the epidemiology of risk factors associated with disseminated cryptococcal disease, and describe a rational workup for a possible acquired immunodeficiency. While rare, 25% of non-AIDS related cryptococcal disease will occur in individuals without an identifiable immunodeficiency and should prompt a work up for cell-mediated immunodeficiency and monitored for closely for progression of other opportunistic infections.
- Published
- 2015
46. Corrigendum to "Somatostatin Negatively Regulates Parasite Burden and Granulomatous Responses in Cysticercosis".
- Author
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Khumbatta M, Firozgary B, Tweardy DJ, Weinstock J, Firozgary G, Bhatena Z, Bulsara T, Siller R, and Robinson P
- Published
- 2015
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47. Contribution of chaperones to STAT pathway signaling.
- Author
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Bocchini CE, Kasembeli MM, Roh SH, and Tweardy DJ
- Abstract
Aberrant STAT signaling is associated with the development and progression of many cancers and immune related diseases. Recent findings demonstrate that proteostasis modulators under clinical investigation for cancer therapy have a significant impact on STAT signaling, which may be critical for mediating their anti-cancer effects. Chaperones are critical for protein folding, stability and function and, thus, play an essential role in the maintenance of proteostasis. In this review we discuss the role of chaperones in STAT and tyrosine kinase (TK) protein folding, modulation of STAT and TK activity, and degradation of TKs. We highlight the important role of chaperones in STAT signaling, and how this knowledge has provided a framework for the development of new therapeutic avenues of targeting STAT signaling related pathologies.
- Published
- 2014
- Full Text
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48. STAT3 signaling is activated preferentially in tumor-initiating cells in claudin-low models of human breast cancer.
- Author
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Wei W, Tweardy DJ, Zhang M, Zhang X, Landua J, Petrovic I, Bu W, Roarty K, Hilsenbeck SG, Rosen JM, and Lewis MT
- Subjects
- Animals, Carcinogenesis pathology, Cell Line, Tumor, Female, Fluorescence, Genes, Reporter, Green Fluorescent Proteins metabolism, Humans, Lentivirus metabolism, Mice, SCID, Models, Biological, Reproducibility of Results, Spheroids, Cellular metabolism, Spheroids, Cellular pathology, Xenograft Model Antitumor Assays, Breast Neoplasms metabolism, Breast Neoplasms pathology, Claudins metabolism, Neoplastic Stem Cells metabolism, Neoplastic Stem Cells pathology, STAT3 Transcription Factor metabolism, Signal Transduction
- Abstract
In breast cancer, a subset of tumor-initiating cells (TIC) or "cancer stem cells" are thought to be responsible for tumor maintenance, treatment resistance, and disease recurrence. While current breast cancer stem cell markers (e.g., CD44(high) /CD24(low/neg) , ALDH positive) have allowed enrichment for such cells, they are not universally expressed and may actually identify distinct TIC subpopulations in the same tumor. Thus, additional markers of functional stem cells are needed. The STAT3 pathway is a critical regulator of the function of normal stem cells, and evidence is accumulating for its important role in breast cancer stem cells. However, due to the lack of a method for separating live cells based on their level of STAT3 activity, it remains unknown whether STAT3 functions in the cancer stem cells themselves, or in surrounding niche cells, or in both. To approach this question, we constructed a series of lentiviral fluorescent (enhanced green fluorescent protein, EGFP) reporters that enabled flow cytometric enrichment of cells differing in STAT3-mediated transcriptional activity, as well as in vivo/in situ localization of STAT3 responsive cells. Using in vivo claudin-low cell line xenograft models of human breast cancer, we found that STAT3 signaling reporter activity (EGFP(+) ) is associated with a subpopulation of cancer cells enriched for mammosphere-forming efficiency, as well as TIC function in limiting dilution transplantation assays compared to negative or unsorted populations. Our results support STAT3 signaling activity as another functional marker for human breast cancer stem cells thus making it an attractive therapeutic target for stem-cell-directed therapy in some breast cancer subtypes., (© AlphaMed Press.)
- Published
- 2014
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49. Monoclonal Antibodies Specific for STAT3β Reveal Its Contribution to Constitutive STAT3 Phosphorylation in Breast Cancer.
- Author
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Bharadwaj U, Kasembeli MM, Eckols TK, Kolosov M, Lang P, Christensen K, Edwards DP, and Tweardy DJ
- Abstract
Since its discovery in mice and humans 19 years ago, the contribution of alternatively spliced Stat3, Stat3β, to the overall functions of Stat3 has been controversial. Tyrosine-phosphorylated (p) Stat3β homodimers are more stable, bind DNA more avidly, are less susceptible to dephosphorylation, and exhibit distinct intracellular dynamics, most notably markedly prolonged nuclear retention, compared to pStat3α homodimers. Overexpression of one or the other isoform in cell lines demonstrated that Stat3β acted as a dominant-negative of Stat3α in transformation assays; however, studies with mouse strains deficient in one or the other isoform indicated distinct contributions of Stat3 isoforms to inflammation. Current immunological reagents cannot differentiate Stat3β proteins derived from alternative splicing vs. proteolytic cleavage of Stat3α. We developed monoclonal antibodies that recognize the 7 C-terminal amino acids unique to Stat3β (CT7) and do not cross-react with Stat3α. Immunoblotting studies revealed that levels of Stat3β protein, but not Stat3α, in breast cancer cell lines positively correlated with overall pStat3 levels, suggesting that Stat3β may contribute to constitutive Stat3 activation in this tumor system. The ability to unambiguously discriminate splice alternative Stat3β from proteolytic Stat3β and Stat3α will provide new insights into the contribution of Stat3β vs. Stat3α to oncogenesis, as well as other biological and pathological processes.
- Published
- 2014
- Full Text
- View/download PDF
50. Chaperonin-containing TCP-1 complex directly binds to the cytoplasmic domain of the LOX-1 receptor.
- Author
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Bakthavatsalam D, Soung RH, Tweardy DJ, Chiu W, Dixon RA, and Woodside DG
- Subjects
- Amino Acid Sequence, Animals, Cattle, Chaperonin Containing TCP-1 genetics, Gene Knockdown Techniques, Human Umbilical Vein Endothelial Cells, Humans, Ligands, Lipoproteins, LDL metabolism, Molecular Sequence Data, Multiprotein Complexes chemistry, Multiprotein Complexes metabolism, Protein Interaction Domains and Motifs, Protein Subunits, RNA, Small Interfering genetics, Scavenger Receptors, Class E genetics, Chaperonin Containing TCP-1 chemistry, Chaperonin Containing TCP-1 metabolism, Scavenger Receptors, Class E chemistry, Scavenger Receptors, Class E metabolism
- Abstract
Lectin-like oxidized low-density lipoprotein receptor (LOX-1) is a scavenger receptor that binds oxidized low-density lipoprotein (OxLDL) and has a role in atherosclerosis development. The N-terminus intracellular region (cytoplasmic domain) of LOX-1 mediates receptor internalization and trafficking, potentially through intracellular protein interactions. Using affinity isolation, we identified 6 of the 8 components of the chaperonin-containing TCP-1 (CCT) complex bound to LOX-1 cytoplasmic domain, which we verified by coimmunoprecipitation and immunostaining in human umbilical vein endothelial cells. We found that the interaction between CCT and LOX-1 is direct and ATP-dependent and that OxLDL suppressed this interaction. Understanding the association between LOX-1 and the CCT complex may facilitate the design of novel therapies for cardiovascular disease., (Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
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